The tight association of phosphate with a molecule, e.g., a protein, has been known since the late nineteenth century. Since then, a variety of covalent linkages of phosphate to proteins have been found. The most common involve esterification of phosphate to serine, threonine, and tyrosine with smaller amounts being linked to lysine, arginine, histidine, aspartic acid, glutamic acid, and cysteine. The occurrence of phosphorylated molecules, e.g., proteins, implies the existence of one or more kinases, e.g., protein kinases, capable of phosphorylating various molecules, e.g., amino acid residues on proteins, and also of phosphatases, e.g., protein phosphatases, capable of hydrolyzing various phosphorylated molecules, e.g., phosphorylated amino acid residues on proteins.
Protein kinases play critical roles in the regulation of biochemical and morphological changes associated with cellular growth and division (D""Urso et al. (1990) Science 250:786-791; Birchmeier et al. (1993) Bioessays 15:185-189). For example, these kinases have been shown to participate in the transmission of signals from growth-factor receptors (Sturgill et al. (1988) Nature 344:715-718; Gomez et al. (1991) Nature 353:170-173), control of entry of cells into mitosis (Nurse (1990) Nature 344:503-508; Mailer (1991) Curr. Opin. Cell Biol. 3:269-275), and regulation of actin bundling (Husain-Chishti et al. (1988) Nature 334:718-721). Protein kinases serve as growth factor receptors and signal transducers and have been implicated in cellular transformation and malignancy (Hunter et al. (1992) Cell 70:375-387; Posada et al. (1992) Mol. Biol. Cell 3:583-592; Hunter et al. (1994) Cell 79:573-582). Alterations in kinase genes and their products can lead to deregulated cell proliferation, a hallmark of cancer. Modulation of these genes and their regulatory activities may permit the control of tumor cell proliferation and invasion.
Protein kinases can be divided into different groups based on either amino acid sequence similarity or specificity for either serine/threonine or tyrosine residues. A small number of dual-specificity kinases have also been described. Within the broad classification, kinases can be further subdivided into families whose members share a higher degree of catalytic domain amino acid sequence identity and also have similar biochemical properties. Most protein kinase family members also share structural features outside the kinase domain that reflect their particular cellular roles. These include regulatory domains that control kinase activity or interaction with other proteins (Hanks et al. (1988) Science 241:42-52).
The present invention is based, in part, on the discovery of a novel kinase family member, referred to herein as xe2x80x9c14815xe2x80x9d. The kinase molecule of the invention shares characteristics with members of the tyrosine kinase family. The nucleotide sequence of a cDNA encoding 14815 is shown in SEQ ID NO:1, and the amino acid sequence of a 14815 polypeptide is shown in SEQ ID NO:2. In addition, the nucleotide sequence of the coding region is depicted in SEQ ID NO:3.
Accordingly, in one aspect, the invention features a nucleic acid molecule which encodes a 14815 protein or polypeptide, e.g., a biologically active portion of the 14815 protein. In a preferred embodiment, the isolated nucleic acid molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO:2. In other embodiments, the invention provides isolated 14815 nucleic acid molecules having the nucleotide sequence shown in SEQ ID NO:1 or SEQ ID NO:3. In still other embodiments, the invention provides nucleic acid molecules that are substantially identical (e.g., naturally occurring allelic variants) to the nucleotide sequence shown in SEQ ID NO:1 or SEQ ID NO:3. In other embodiments, the invention provides a nucleic acid molecule which hybridizes under a stringent hybridization condition as described herein to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO:3, wherein the nucleic acid encodes a full length 14815 protein or an active fragment thereof.
In a related aspect, the invention further provides nucleic acid constructs which include a 14815 nucleic acid molecule described herein. In certain embodiments, the nucleic acid molecules of the invention are operatively linked to native or heterologous regulatory sequences. Also included are vectors and host cells containing the 14815 nucleic acid molecules of the invention e.g., vectors and host cells suitable for producing polypeptides.
In another related aspect, the invention provides nucleic acid fragments suitable as primers or hybridization probes for the detection of 14815-encoding nucleic acids.
In still another related aspect, isolated nucleic acid molecules that are antisense to a 14815 encoding nucleic acid molecule are provided.
In another aspect, the invention features 14815 polypeptides, and biologically active or antigenic fragments thereof that are useful, e.g., as reagents or targets in assays applicable to treatment and diagnosis of kinase-associated or other 14815-associated disorders. In another embodiment, the invention provides 14815 polypeptides having a 14815 activity. Preferred polypeptides are 14815 proteins including at least one protein kinase domain, at least one, two, three, preferably four armadillo/beta-catenin-like repeat domains, and, preferably, having a 14815 activity, e.g., a 14815 activity as described herein.
In other embodiments, the invention provides 14815 polypeptides, e.g., a 14815 polypeptide having the amino acid sequence shown in SEQ ID NO:2; an amino acid sequence that is substantially identical to the amino acid sequence shown in SEQ ID NO:2; or an amino acid sequence encoded by a acid nucleic molecule having a nucleotide sequence which hybridizes under a stringent hybridization condition as described herein to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:3, wherein the nucleic acid encodes a full length 14815 protein or an active fragment thereof.
In a related aspect, the invention further provides nucleic acid constructs which include a 14815 nucleic acid molecule described herein.
In a related aspect, the invention provides 14815 polypeptides or fragments operatively linked to non-14815 polypeptides to form fusion proteins.
In another aspect, the invention features antibodies and antigen-binding fragments thereof, that react with, or more preferably specifically or selectively bind 14815 polypeptides.
In another aspect, the invention provides methods of screening for compounds that modulate the expression or activity of the 14815 polypeptides or nucleic acids.
In still another aspect, the invention provides a process for modulating 14815 polypeptide or nucleic acid expression or activity, e.g., using the compounds identified in the screens described herein. In certain embodiments, the methods involve treatment of conditions related to aberrant activity or expression of the 14815 polypeptides or nucleic acids, such as conditions or disorders involving aberrant or deficient kinase function or expression. Examples of such disorders include, but are not limited to, cellular proliferative and/or differentiative disorders, neurological disorders, inflammatory disorders, liver disorders, apoptotic disorders, metabolic disorders and hormonal disorders.
The invention also provides assays for determining the activity of or the presence or absence of 14815 polypeptides or nucleic acid molecules in a biological sample, including for disease diagnosis.
In a further aspect, the invention provides assays for determining the presence or absence of a genetic alteration in a 14815 polypeptide or nucleic acid molecule, including for disease diagnosis.
In another aspect, the invention features a two dimensional array having a plurality of addresses, each address of the plurality being positionally distinguishable from each other address of the plurality, and each address of the plurality having a unique capture probe, e.g., a nucleic acid or peptide sequence. At least one address of the plurality has a capture probe that recognizes a 14815 molecule. In one embodiment, the capture probe is a nucleic acid, e.g., a probe complementary to a 14815 nucleic acid sequence. In another embodiment, the capture probe is a polypeptide, e.g., an antibody specific for 14815 polypeptides. Also featured is a method of analyzing a sample by contacting the sample to the aforementioned array and detecting binding of the sample to the array.
Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.